Background: \r\n', u'Rotavirus is the major important cause of infectious infantile gastroenteritis and diarrhea, especially the children aged 6 months to 24 months. Some efforts made to provide safe water and cultivate the hygiene have not reduced the diarrhea caused by rotaviruses. Develope a vaccine agaisnt this virus is of high importance. A question about the production of this kind of vaccine targeted athow to make human rotaviruses strains adapt cultured cells.\r\n', u'Objectives\r\n', u'To determine the multiplicity of infection (MOI) of human rotaviruses strains in vero cell.\r\n', u'Subjects and method: \r\n', u'The multiplicity of infection of Human rotaviruses strains (G1 P8, G1P4, and G4P6) were selected and the CDC- Atlanta US supplied the vero cell.Use vero cells cultivate, temper the vero cell and, applying direct immunofluorescence to determine the multiplicity of G1P8, G1P4, C4P6.\r\n', u'Results:\r\n', u'The MOI of G1P8, G1P4 and C4P6 are 0.02 ffu/cell, 0.15ffu/cell, and 0.35 ffu/cell, respectively.\r\n', u'Conclusion:\r\n', u'Using the vero cell to multiple the rotaviruses is the effective part during the production of vaccines against rotaviruses. The MOI of human rotaviruses strains G1P8, G1P4 and G4P6 are in order:0,02 ffu/cell, 0,15ffu/cell, and 0,35% ffu/cell. \r\n', u'
Rotavirus ; Vero Cells ; Cell Culture Techniques ;

Introduction: Rabies is a serious problem in the area of public health in developing countries including Vietnam. The death rate is almost 100%, however rabies can be prevented and preventively treated by vaccine, or a combination between vaccine with rabies resistant serum. The production of Vero cell culture vaccine is becoming a common trend worldwide because of its effective protection and safety. There is a requirement for the domestic production of cell cultured rabies vaccine. \r\n', u'Objectives: To determine the adaptive ability of VNUCOVO - 32 rabies vaccine strain into Vero cell. \r\n', u'Subjects and Method: Rabies vaccine propagation strains the rat\u2019s kidney cells. VNUCOVO - 32 was cultured into Vero cell with different conditions such as pH, temperature, multiplicity of infection (MOl) and time of virus harvests to discover the optimal conditions for virus propagation. \r\n', u'Results: The optimal condition for VNUCOVO - 32 propagation into vero cell with MOl 0.3, pH 7.4 and temperature is 37\xb0C. The highest titer achieves 103,4FFLJ/ml. The best time for virus harvest is 12 - 13 days post inoculation. \r\n', u'Conclusion: VNUCOVO rabies vaccine strain can penetrate and propagate into vera cell. \r\n', u'
Rabies virus ; VNKONO \u2013 32 strain ; Vero cell

Recovery from potentially lethal damage (PLDR) after irradiation was studied in plateau-phase culture of Vero cells in vitro. Unfed plateau-phase cells were irradiated with dose of 1 to 9 gy using Cs-137 irradiator. Cells then were incubated again and left in situ for 0, 1, 2, 3, 4, 5, 6 and 24 hours and then were trypsinized, explanted, and subcultured in fresh RPMI-1640 media containing 0.33% agar. Cell survival was measured by colony forming ability. An adequate number of heavily irradiated Vero cells were added as feeder cells to make the total cell number constant in every culture dish. As the postirradiation in situ incubation time increased, surviving fraction increased saturation level at 2 to 4 hours after in situ incubation. As the radiation dose increased, the rate of PLDR also increased. In analysis of cell survival curve fitted to the linear-quadratic model, the linear inactivation coefficient (alpha) decreased largely and reached nearly to zero but the quadratic inactivation coefficient (beta) increased minimally by increment of postirradiation in situ incubation time. So PLDR mainly affected the damage expressed as alpha. In the multitarget model, significant change was not obtained in D0 but in Dq. Therefore, shoulder region in cell survival curve was mainly affected by PLDR and terminal slope was not influenced at all. And dose-modifying factor by PLDR was relatively higher in shoulder region, that is, in low dose area below 3 gy.
Agar ; Cell Count ; Cell Survival ; Feeder Cells ; Shoulder ; Trypsin ; Vero Cells*

BACKGROUND: Cell culture is the golden standard method for Herpes simplex virus (HSV) isolation. However, some specimens require many days to develop any cytopathic effect (CPE). We developeda rapid sensitive culture technique for HSV isolations. METHODS: This study included a total of 133 patients with suspected HSV infection. Specimens were centrifuged onto a Vero cell monolayer in a shell vial. The CPE was observed daily during the5-day incubation by inverted-phase microscope. The direct immunofluorescence (DIF) stain with aHSV specific antibody was performed 2 days after sample inoculation. The negative samples in theDIF stain were reinoculated in the new shell vials after extraction of the monolayer. Polymerase chainreaction for HSV detection was performed using the original samples. RESULTS: The CPE was observed 30 (64%), 39 (83%), 43 (92%), 44 (94%), and 46 (98%) cases at1, 2, 3, 4, and 5 days incubation, respectively. The DIF stain detected 46 cases (98%) at 2 days incubation. The CPE was observed in another 7 cases at 1-day incubation after the reinoculation of negative samples. The PCR detected 47 (100%) of 133 cases. CONCLUSIONS: The reinoculation of negative sample in a shell vial culture is a rapid sensitive methodfor HSV isolation.
Cell Culture Techniques ; Culture Techniques ; Fluorescent Antibody Technique, Direct ; Humans ; Polymerase Chain Reaction ; Simplexvirus* ; Vero Cells

cell culture processes for viral vaccine production are mainly based on adherent cell culture systems using serum, which are associated with expensive and labor-intensive processes to produce large amounts of viral vaccine strains. In this study, we investigated whether Vero cells could be grown in serum-free and shaking suspension conditions. Furthermore, we assessed the ability of the Vero cell suspension culture system to produce adenovirus type 5 (Ad5), compared to that of the adhesive Vero cell culture system.MATERIALS AND METHODS: We tested the feasibility of commercial serum-free media for Vero cell culture. For the adaptation of Vero cells in suspension culture, adhesive Vero cells were added in the early phase of shaking suspension culture, and 50 days after shaking suspension culture, suspension-adapted Vero cells were subcultured continuously. To assess the virus production ability of Vero cells in suspension, the cells were infected with Ad5-green fluorescent protein and evaluated based on their fluorescence intensity.RESULTS: The Vero cells grown in OptiPRO serum-free medium showed no changes in morphology and growth rate, but MRC-5 and FRhk-4 cells showed morphological changes and decreased growth rate, respectively. The Vero cells were well adapted to the suspension culture system. The Vero cells in suspension showed a better Ad5 production ability than the adherent Vero cells.CONCLUSION: Vero cells can be grown in OptiPRO serum-free medium. Further, our suspension culture-adapted Vero cells may be suitable to produce viral vaccine strains due to their high ability to produce viruses such as Ad5.]]>
Adenoviridae ; Adhesives ; Cell Culture Techniques ; Culture Media, Serum-Free ; Fluorescence ; Vero Cells

OBJECTIVETo find a sensitive cytotoxic response to reflect the bio-toxicity of trace organic pollutants, the sensitivity and reliability of morphological change and proliferation inhibition of Vero cells exposed to 2, 4, 6-trichlorophenol (TCP) and the leachate from products related to drinking water (PRDW) were compared, and the mechanism of the morphological change in Vero cells exposed to chemical pollutants was studied.

METHODSVero cells were treated by different concentration of TCP and the leachate from PRDW. Methylthiazol-2-yl-2, 5-diphenyl tetrazolium bromide (MTT) assay was carried out for proliferation inhibition. Bioluminescence method was carried out as another method to test the toxicity of TCP. Flow Cytometry assay was used to test cell Apoptosis and damage of cell-membrane.

RESULTS0.25 mg/L TCP had an effect on cell morphology, and the proportion of morphologically changed cells increased with increasing TCP concentration. At low TCP concentrations, inhibition of cell proliferation did not seem to correlate to TCP concentration, and was negative when TCP concentration was <1.0 mg/L. After exposure to leachate from PRDW extracted at different temperatures, the percentage of morphologically changed cells increased with extracting temperature, but the inhibition of cell proliferation failed to reflect the correlation between extracting temperature and proliferation inhibition of Vero cells. Although the Sensitivity of bioluminescence method seems to be similar to morphological change in Vero cells, the bacterial in this method is not homologous enough with human body cells to reflect the toxicity to human body. These imply cell morphological change is a more sensitive and reliable method to reflect bio-toxicity of organic pollutants than proliferation inhibition. Flow cytometry analysis and cell rejuvenation experiments indicated cell membrane damage, which results in cell morphological change, was an early and sensitive cytotoxic response comparing with necrosis.

CONCLUSIONThese results indicated that the cell membrane toxicity represented by morphological changes is a more sensitive and reliable method to indicate the composite bio-toxicity of trace chemicals than proliferation inhibition, inhibition on bioluminescence and necrosis. Nevertheless, the quantification of morphological change should be studied further.

Animals ; Cell Division ; drug effects ; Cell Survival ; drug effects ; Cercopithecus aethiops ; Vero Cells ; Water Pollutants, Chemical ; toxicity

OBJECTIVETo investigate the differentiation of human testicular spermatogenic cells during in vitro culture.

METHODSTesticular cells of obstructive azoospermic patients' testis biopsies were dispersed employing mechanic methods. Then, (1) mixed testicular cells were applied to in vitro culture, and changes of the ratio of elongating spermatids and all round cells were analyzed during mixed cell culture; (2) round spermatids were picked up from the mixed cells employing micromanipulator, followed by differentiation of the isolated round spermatids during microdrop culture.

RESULTSThe ratio of the elongating spermatids increased significantly (P < 0.05) after 24 hours of mixed cell culture in HTF medium supplemented with FSH and testosterone. During single round spermatid culture, transformation of the round spermatid to elongating spermatid with newly formed flagellum was observed, and the transformation ratio within 48 hours of microdrop culture was 3.54%. The differentiation of human testicular spermatogenic cells cultured in Vero cell conditioned medium was similar to that cultured in HTF medium.

CONCLUSIONHuman testicular round spermatids can differentiate to elongating spermatids during in vitro culture. Vero cell conditioned medium does not promote the differentiation of human testicular round spermatids to elongating spermatids.

Animals ; Cell Differentiation ; Cells, Cultured ; Cercopithecus aethiops ; Humans ; Infertility, Male ; pathology ; Male ; Spermatids ; cytology ; Testis ; cytology ; Vero Cells

Cyclin-dependent protein kinase (CDK) plays an important role in the replication of herpes simplex virus (HSV) and other important human disease viruses. But which kinds of CDK are required in the replication of HSV is still not clear. In this study, we infected dominant negative CDK2 cell line with different multiplicity of infection (MOI) of HSV-1-KOS strain (abbreviated as HSV below), and the results showed that the yield of HSV depended on the MOI; the replication of HSV delayed about 3 h as compared with that of the control in the one-step growth curve replication experiments; the CDK2 activity was induced 6 h post HSV infection and reached the highest 9 h post infection; the HSV went into rapid productive replication after the CDK2 was induced. We propose herein that the CDK2 is required in the initiation of replication of HSV.
Animals ; Cell Proliferation ; Cercopithecus aethiops ; Cyclin-Dependent Kinase 2 ; physiology ; HT29 Cells ; Humans ; Simplexvirus ; physiology ; Vero Cells ; Virus Replication

OBJECTIVETo research the cytotoxicity and in vitro antiproliferative effect of the six flavone compounds extracted from Laggera pterodonta.

METHODThe cytotoxicity on the normal cells and antiproliferative effect on tumor cells were tested by MTT assay, and then the preliminary structure-activity relationship was analysed. The phase distribution of the cell cycle and apoptosis rate were analyzed by flow cytometry.

RESULTThe results of MTT assay showed 5,7,3',4'-tetramethoxy-3-hydroxyflavone and chrysosplenetin B inhibited growth of A549 and Hela cells significantly with a dose dependent mode, while exhibited low cytotoxicity to the two normal cells, Vero and EVC304. Both compounds contain ortho-phenolic methoxyl moietys in their structures. Flow cytometry analysis revealed that Hela cells treated with increasing quantities of chrysosplenetin B increased the percentage of cells in the G2/M phase, and Hela and A549 cells treated with increasing quantites of the 5,7,3',4'-tetramethoxy-3-hydroxyflavone and chrysosplenetin B increased the apoptosis rates.

CONCLUSIONThe 5,7,3',4'-tetramethoxy-3-hydroxyflavone and chrysosplenetin B extracted from L. pterodonta showed high antiproliferative effect on cancer cells with low cytotoxicity on normal cells, and took the effects on A549 and Hela cells through the hold-up of the G2/M phase of the cell cycle and induction of the apoptosis.

Animals ; Asteraceae ; chemistry ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cercopithecus aethiops ; Flavones ; chemistry ; pharmacology ; Flavonoids ; pharmacology ; Flow Cytometry ; HeLa Cells ; Humans ; Vero Cells

OBJECTIVETo develop a novel rapid protocol for the detection of occult disseminated tumor cells by a recombinant herpes simplex virus expressing GFP (HSV(GFP)).

METHODSTumor cells of seven cell lines were exposed to HSV(GFP) and then examined for GFP expression by fluorescence microscopy. Various numbers of tumor cells (10, 100, 1000, 10 000) were mixed into 2 ml human whole blood, separated with lymphocytes separation medium, exposed to HSV(GFP), incubated at 37°C for 6 - 24 h and then counted for the number of green cells under the fluorescence microscope. Some clinical samples including peripheral blood, pleural effusion, ascites, spinal fluid from tumor-bearing patients were screened using this protocol in parallel with routine cytological examination.

RESULTSHSV(GFP) was able to infect all 7 tumor cell lines indicating that the HSV(GFP) can be used to detect different types of tumor cells. The detection sensitivity was 10 cancer cells in 2 ml whole blood. In the clinical samples, there were 4/15 positive by routine cytological examination but 11/15 positive by HSV(GFP), indicating a higher sensitivity of this new protocol.

CONCLUSIONRecombinant herpes simplex virus-mediated green fluorescence is a simple and sensitive technique for the identification of occult disseminated cancer cells including circulating tumor cells (CTCs).

Animals ; Cell Line, Tumor ; Cercopithecus aethiops ; Green Fluorescent Proteins ; metabolism ; Humans ; Neoplastic Cells, Circulating ; metabolism ; pathology ; Recombinant Proteins ; metabolism ; Sensitivity and Specificity ; Simplexvirus ; metabolism ; Vero Cells